StCDPK5 confers resistance to late blight pathogen but increases susceptibility to early blight pathogen in potato via reactive oxygen species burst
StCDPK5 confers resistance to late blight pathogen but increases susceptibility to early blight pathogen in potato via reactive oxygen species burst
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DOI：
10.1111/j.1469-8137.2012.04226.x
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发表时间：
2012-10-01
期刊：
影响因子：
9.4
通讯作者：
Yoshioka, Hirofumi
中科院分区：
文献类型：
--
作者：
Kobayashi, Michie;Yoshioka, Miki;Yoshioka, Hirofumi
马铃薯（Solanum tuberosum）钙依赖性蛋白激酶（StCDPK5）可磷酸化质膜RBOH（呼吸爆发氧化酶同源物）蛋白的n端区域，并参与strbohb介导的活性氧（ROS）爆发。本构活性形式StCDPK5VK为防御反应中RBOH的功能获得分析提供了有用的工具。StCDPK5-和StCDPK5VK-绿色荧光蛋白融合蛋白主要靶向质膜，StCDPK5VK条件表达激活StRBOHA-D。通过双分子荧光互补实验证实了相互作用。我们在病原菌诱导启动子的控制下，培育了含有StCDPK5VK的转基因马铃薯植株，以研究ROS爆发对枯萎病病原菌防御反应的作用。晚疫病病原菌疫霉（Phytophthora infestans）和早疫病病原菌茄疫病菌（Alternaria solani）的毒力分离物在转基因植株侵染部位诱导了超敏反应样细胞死亡，并伴随ROS的产生。转基因植物显示出对近专性半生物营养病原菌P. infestans的抗性，相比之下，对坏死性病原菌A. solani的敏感性增加。这些结果表明，rboh依赖性ROS有助于对近专性病原体的基础防御，但在抵抗中起负作用，或在坏死性病原体引起的疾病病变扩大中起积极作用。
Potato (Solanum tuberosum) calcium-dependent protein kinase (StCDPK5) has been shown to phosphorylate the N-terminal region of plasma membrane RBOH (respiratory burst oxidase homolog) proteins, and participate in StRBOHB-mediated reactive oxygen species (ROS) burst. The constitutively active form, StCDPK5VK, provides a useful tool for gain-of-function analysis of RBOH in defense responses. StCDPK5- and StCDPK5VK-green fluorescent protein fusion proteins were predominantly targeted to the plasma membrane, and conditional expression of StCDPK5VK activated StRBOHA-D. The interaction was confirmed by bimolecular fluorescence complementation assay. We generated transgenic potato plants containing StCDPK5VK under the control of a pathogen-inducible promoter to investigate the role of ROS burst on defense responses to blight pathogens. Virulent isolates of the late blight pathogen Phytophthora infestans and the early blight pathogen Alternaria solani induced hypersensitive response-like cell death accompanied by ROS production at the infection sites of transgenic plants. Transgenic plants showed resistance to the near-obligate hemibiotrophic pathogen P. infestans and, by contrast, increased susceptibility to the necrotrophic pathogen A. solani. These results indicate that RBOH-dependent ROS contribute to basal defense against near-obligate pathogens, but have a negative role in resistance or have a positive role in expansion of disease lesions caused by necrotrophic pathogens.